Title

Author

Defense Date

2009

Document Type

Thesis

Degree Name

Master of Science

Department

Physiology

First Advisor

Louis Dr. De Felice

Abstract

Porphyromonas gingivalis, a gram-negative anaerobic bacterium, is implicated as a major etiological agent in the initiation and progression of severe forms of periodontal disease. It has been reported that gingivitis and periodontal disease affect roughly 50.3% and 35% of the adult population, respectively. Therefore, approximately over 85% of the adult population may suffer from some form of gingival disease. Porphyromonas gingivalis, an established periodontopathogen, requires hemin for growth. Although multiple hemin uptake systems appear to be present in this organism, their specific role in hemin uptake and virulence remains unknown. Pigmentation is thought to result from the accumulation of iron protoporphyrin IX (FePPIX) derived from erythrocyte hemoglobin. It has been observed that mutations abolishing activity of the Lys-X specific cysteine protease, Kgp, resulted in loss of black pigmentation of P. gingivalis W83; they were less virulent than their wild-type counterparts. Thus, we have observed that Kgp degradation of fibrinogen deregulates the clotting cascade, thereby minimizing the availability of free erythrocytes. Additionally, Kgp binds erythrocytes and degrades them, releasing hemoglobin. The interference with mechanisms involved in the accumulation of black pigmentation may be significant in controlling the pathogenic potential of P. gingivalis. These results suggest that Lys-gingipain protease is a principal protein involved in acquisition of hemin from hemoglobin as well as a major factor in transport, by affecting the accumulation of FePPIX on the bacterial cell surface. Microarray analysis indicates a change in the expression of key enzymes and proteins required for hemin uptake, iron storage, electron transport and oxidative stress. Therefore, interference with mechanisms involved in accumulation of black pigmentation may be significant in controlling the pathogenic potential of P. gingivalis.